• Title/Summary/Keyword: Isolation Enhancement

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Bioactivity of Metabolites from Actinomycetes Isolates from Red Sea, Egypt

  • Osman, Mohamed E.;El-nasr, Amany A. Abo;Hussein, Hagar M;Hamed, Moaz M
    • Microbiology and Biotechnology Letters
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    • v.50 no.2
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    • pp.255-269
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    • 2022
  • Actinomycetes isolated from marine habitats represent a promising source of bioactive substances. Here, we report on the isolation, identification, productivity enhancement and application of the bioactive compounds of Streptomyces qinglanensis H4. Eighteen marine actinomycetes were isolated and tested for resistance to seven bacterial diseases. Using 16S rRNA sequencing analysis (GenBank accession number MW563772), the most powerful isolate was identified as S. qinglanensis. Although the strain produced active compound(s) against a number of Gram-negative and Gram-positive bacteria, it failed to inhibit pathogenic fungi. The obtained inhibition zones were 22.0 ± 1.5, 20.0 ± 1, 16.0 ± 1, 12.0 ± 1, 22.0 ± 1 and 24.0 ± 1 mm against Bacillus subtilis ATCC 6633, Escherichia coli ATCC 19404, Enterococcus faecalis ATCC 29212, Pseudomonas aeruginosa ATCC 9027, Candida albicans ATCC 10231 and Staphylococcus aureus ATCC6538, respectively. To maximize bioactive compound synthesis, the Plackett-Burman design was used. The productivity increased up to 0.93-fold, when S. qinglanensis was grown in optimized medium composed of: (g/l) starch 30; KNO3 0.5; K2HPO4 0.25; MgSO4 0.25; FeSO4·7H2O, 0.01; sea water concentration (%) 100; pH 8.0, and an incubation period of 9 days. Moreover, the anticancer activity of S. qinglanensis was tested against two different cell lines: HepG2 and CACO. The inhibition activities were 42.96 and 57.14%, respectively. Our findings suggest that the marine S. qinglanensis strain, which grows well on tailored medium, might be a source of bioactive substances for healthcare companies.

Development of High-performance Microwave Water Surface Current Meter for General Use to Extend the Applicable Velocity Range of Microwave Water Surface Current Meter on River Discharge Measurements (전자파표면유속계를 이용한 하천유량측정의 적용범위 확장을 위한 고성능 범용 전자파표면유속계의 개발)

  • Kim, Youngsung;Won, Nam-Il;Noh, Joonwoo;Park, Won-Cheol
    • Journal of Korea Water Resources Association
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    • v.48 no.8
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    • pp.613-623
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    • 2015
  • To overcome the difficulties of discharge measurements during flood season, MWSCM(micowave water surface current meter) which measures river surface velocities without contacting water has been applied in field work since its development. The existing version of MWSCM is for floods so that its applicability is low due to the short periods of floods. Therefore the renovative redesign of MWSCM to increase the applicability was conducted so that it can be applied to the discharge measurements during normal flows as well as flood ones by extending the measurable range of velocity. A newly developed high-performance MWSCM for general use can measure the velocity range of 0.03-20.0 m/s from flood flows to normal flows, whereas MWSCM for floods can measure the velocity range of 0.5-10.0 m/s. The improvement of antenna isolation between transmitter and receiver to block the inflow of transmitted singals to receiver and the improvement of phase noise of oscillator are necessary for detecting low velocity with MWSCM technology. Separate type antenna of transmitting and receiving signals is developed for isolation enhancement and phase locked loop synthesizer as an oscillator is applied to high-performance MWSCM for general use. Microwave frequency of 24 GHz is applied to the new MWSCM rather than 10 GHz to make the new MWSCM small and light for convenient use of it at fields. Improvement requests on MWSCM for floods-stable velocity measurement, self test, low power consumtion, and waterproof and dampproof-from the users of it has been reflected on the development of the new version of MWSCM.

Comparison among the Sperm Preparation Methods on the Human Spermatozoa (인간정자에 있어서 정자처리법의 비교)

  • Pang, Myung-Geol;Chung, Ku-Min;Shin, Chang-Jae;Kim, Jung-Gu;Moon, Shin-Yong;Chang, Yoon-Seok;Lee, Jin-Yong;Lee, Sang-Hoon;Chung, Yung-Chai;Kim, Chang-Keun
    • Clinical and Experimental Reproductive Medicine
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    • v.20 no.2
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    • pp.107-115
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    • 1993
  • Procedures to separate motile. normal & motile and acrosome-reacted sperm with high efficiency have clinical application in Assisted Reproductive Technology in terms of increasing the probability of fertilization by a normal sperm and subsequent normal embryonic development. This study evaluated the effects of 10 sperm preparation techniques [Swim-up from a washed pellet (SU). Swim-up from semen (SO). Continuous Percoll Gradients I (PIC). Discontinuous Percoll Gradients I (PID). Continuous Percoll Gradients II(P II C). Discontinuous Percoll Gradients II(P II D), SpermPrep (SFC). Wang's tube (WT). Albumin Gradients (AG), Low temperature capacitation (LTC)] on motility (%), normal morphology (%), motile sperm recovery rate(%). morphologically normal & motile sperm recovery rate (%), true acrosome reaction (%) and fertilizing ability. A P II D proved to be an effective means of separating morphologically normal & motile sperm. Our results indicated the P II D has advantages as compared with other methods in terms of recovery rate. enhancement of motility and normal morphology. And a LTC seems to be an effective means of enhancing the true acrosome reaction and fertilizing ability. These results suggest that the combined method of LTC and P II D for separation of morphologically normal & motile sperm and acrosome reacted sperm may be a useful procedure for intrauterine insemination and in vitro fertilization in the management of male factor infertility as well as for isolation of subpopulation of sperm for basic research.

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Isolation and Identification of a New Gene Related to Salt Tolerance in Chinese Cabbage (배추에서 신규 염 저항성 관련 유전자 분리 및 검정)

  • Yu, Jae-Gyeong;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.31 no.6
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    • pp.748-755
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    • 2013
  • This study was conducted to find a salt tolerance gene in Brassica rapa. In order to meet this objective, we analyzed data from a KBGP-24K oligo chip [BrEMD (Brassica rapa EST and microarray database)] of the B. rapa ssp. pekinensis 'Chiifu' under salt stress (250 mM NaCl). From the B. rapa KBGP-24K microarray chip analysis, 202 salt-responsive unigenes were primarily selected under salt stress. Of these, a gene with unknown function but known full-length sequence was chosen to closely investigate the gene function. The selected gene was named BrSSR (B. rapa salt stress resistance). BrSSR contains a 285 bp open reading frame encoding a putative 94-amino acid protein, and a DUF581 domain. The pSL94 vector was designed to over-express BrSSR, and was used to transform tobacco plants for salt tolerance analysis. T1 transgenic tobacco plants that over-expressed BrSSR were selected by PCR and DNA blot analyses. Quantitative real-time RT PCR revealed that the expression of BrSSR in transgenic tobacco plants increased by approximately 3.8-fold. Similar results were obtained by RNA blot analysis. Phenotypic characteristics analysis showed that transgenic tobacco plants with over-expressed BrSSR were more salt-tolerant than the wild type control under 250 mM NaCl for 5 days. Based on these results, we hypothesized that the over-expression of BrSSR may be closely related to the enhancement of salt tolerance.

Isolation of Flavonoids and Lignans from the Stem Wood of Lindera obtusiloba Blume (생강나무(Lindera obtusiloba Blume) 목부로부터 Flavonoid 및 Lignan 화합물의 분리)

  • Seo, Kyeong-Hwa;Baek, Mi-Young;Lee, Dae-Young;Cho, Jin-Gyeong;Kang, Hee-Cheol;Ahn, Eun-Mi;Baek, Nam-In;Lee, Youn-Hyung
    • Journal of Applied Biological Chemistry
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    • v.54 no.3
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    • pp.178-183
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    • 2011
  • The stem woods of Lindera obtusiloba Blume were extracted in 80% aqueous methanol and the concentrated extract was partitioned with ethyl acetate (EtOAc), butanol (n-BuOH), and $H_2O$, successively. From the EtOAc and n-BuOH fractions, five compounds were isolated through the repeated silica gel, octadecyl silica gel, and Sephadex LH-20 column chromatographies. On the basis of spectroscopic data including mass spectrometry, IR, $^1H$-NMR, $^{13}C$-NMR, distortionless enhancement by polarization transfer, and two-dimensional-NMR gradient correlated spectroscopy (gCOSY), gradient heteronuclear single quantum correlation (gHSQC), gradient heteronuclear multiple bonding connectivity (gHMBC), the chemical structures of the compounds were determined as asarinin (1), (+)-catechin (2), (-)-epicatechin (3), hyperin (4), and nudiposide (5). Compounds 1 and 5 were isolated for the first time from the stem wood of L. obtusiloba Blume.

Isolation and Characterization of Phosphate Solubilizing Bacteria Pantoea Species as a Plant Growth Promoting Rhizobacteria (식물 생장 촉진 활성을 가진 인산분해 미생물 Pantoea 종의 분리 및 특성 규명)

  • Yun, Chang Yeon;Cheong, Yong Hwa
    • Journal of Life Science
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    • v.26 no.10
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    • pp.1163-1168
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    • 2016
  • Plant growth-promoting rhizobacteria (PGPR) have gained worldwide importance and acceptance due to their agricultural benefits. These microorganisms are potential tools for sustainable agriculture, with effects on plant growth, biofertilization, induced systemic resistance, and biocontrol of plant pathogens. In this study, four different Pantoea species were isolated from field soil, and their plant growth-promoting characteristics were studied. Based on 16S rDNA gene sequencing analyses, the se were grouped into Pantoea ananatis, Pantoea citrea, Pantoea dispersa, Pantoea vagans and named as Pa1, Pc1, Pd1, Pv1, respectively. All of these strains have their ability for solubilization of insoluble phosphate depending on pH decrease at the range around pH 5 at 1days after inoculation and production of plant hormone indole acetic acid (IAA) with 85.3±16.3 μg/ml of Pa1, 183.9±16.8 μg/ml of Pc1, 28.8±17.3 μg/ml of Pd1 and 114.1±16.5 μg/ml of Pv1, respectively. Pa1, Pc1 and Pd1 also have high activity for production of gibberellin (GA3) hormone with 331.1±19.2 μg/ml of Pa1, 288.5±16.8 μg/ml of Pc1, 309.2±18.2 μg/ml of Pd1, but Pv1 does not. Furthermore, all these species have significantly promoted the growth of the lettuce seedling plants at the range around 32~37% for fresh weight and 10~15% for shoot length enhancement, so that these microbe could be used as a potential bio-fertilizer agents.

A Study on the Resilience Process of Persons with Disabilities (중도장애인의 레질리언스(Resilience) 과정에 관한 연구)

  • Kim, Mi-Ok
    • Korean Journal of Social Welfare
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    • v.60 no.2
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    • pp.99-129
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    • 2008
  • This study analyzed the resilience process of persons with disabilities using the grounded theory approach. To conduct this study, the researcher conducted in-depth interviews with 8 persons with disabilities. In data analysis, this study identified 393 concepts on the resilience process of persons with disabilities and the concepts were categorized into 45 sub-categories and 18 primary categories. In the paradigm model on the resilience process of persons with disabilities, it was identified that casual conditions included 'unawareness of disability before being disability', 'extreme pain', 'repressing psychological pain', and the contingent conditions were 'dis-empowerment by staying in home', 'isolation by himself with difficulty in accepting the disability', 'experience of frustration from social barriers with prejudice against persons with disabilities'. Also, it was identified that the resilience process could be dependent on the type and the degree of the disability, the gender, and the length of time being disability. In spite of the casual and contingent conditions, the central way in which persons with disabilities could acquire resilience was identified as 'enhancement of the power of positive thinking'. The control conditions which accelerate or retard central phenomenon were 'the awareness of not being alone through family, friends, neighborhood and the social system' externally and 'finding purpose in life through religion and help from other persons with disabilities', internally. The action/interactional sequences enhanced the efforts, self searching and active acting, and as a result, persons with disabilities could find comfort in life, participate in society and change the perspective of disability in society. The core categories of resilience process in persons with disabilities were a belief in affirmation and choice of life by initiative. In the process analysis, stages developed in the following: 'pain', 'strangeness', 'reflection', 'daily life'. This stage was more continuous and causal than discrete and complete. In this process, the types of resilience of persons with disabilities are divided into 'existence reflection', 'course development', 'implicit endeavor', and 'active execution'. This study showed the details of the paradigm models, the process and types with an in-depth understanding of the resilience process of persons with disabilities using grounded theory as well as theory construction and policy and clinical involvement on the study of persons with disabilities.

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Strain Improvement through Protoplast Formation and Mutation of Inonotus obliquus Mycelia for Enhanced Production of Innerpolysaccharides (IPS) in Suspended Mycelial Cultures (Inonotus obliquus 의 균사체 액상배양에서 원형질체 형성과 돌연변이를 통한 단백다당체 고생산성 균주 개발)

  • Hong, Hyeong-Pyo;Jeong, Yong-Seob;Chun, Gie-Taek
    • KSBB Journal
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    • v.25 no.2
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    • pp.155-166
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    • 2010
  • Studies on the production of cell-wall bound innerpolysaccharides (IPS) (soluble ${\beta}$-D-glucan) have been performed by use of suspended myelial cultures of Inonotus obliquus. This product has promising potentials as an effective antidiabetic as well as an immunostimulating agents. As a first step to enhanced production of IPS, Intensive strain improvement programs were carried out by obtaining a large amounts of protoplasts for the isolation of single cell colonies. Rapid and large screening of high-yielding producers was possible because about fivefold higher amount of protoplasts ($2.3{\times}10^6$ protoplasts/mL) could be recovered with relatively high regeneration rates of $10^{-2}{\sim}10^{-3}$ by applying a modified filtration method, as compared to the previously used trapping method. A basic protocol necessary for UV-mutation of the protoplasts was also developed, resulting in several overproducing variants with good fermentation properties. Since the amount of IPS extracted from the mycelial cell walls of I. obliquus turned out to be almost constant per g DCW, increase in cell mass was considered the most important factor for the enhancement in IPS production. Therefore, attempts were made to screen mutant cells showing rapid mycelial growth rate in the final suspended cultures. Notably, the mutant strains showing an active cellgrowth in the preceding solid growth cultures were observed to produce higher amount of IPS in the suspended fermentations as well. A striking mutant, OBLQ756-15-5 strain, obtained from the survivors of a harsh UV-treated condition (97% death rate) was found to stably produce as high cell mass as 22 g DCW/L in the final fermentations. Currently, this strain is being tested for development of a scaled-up fermentation process for mass production of IPS.